Journal of Peking University(Health Sciences) 2001;33(2):167-169

The expression and purificiation of human Annexin Ⅴ overexpressed in E. coli and the detection of apoptosis

Chunhui DI ; Qun SHI ; Yingmei ZHANG ; Cheng ZHAO ; Yingcheng ZHONG ; Dawu HUANG ; Dalong MA

Country

China

Language

Chinese

Abstract

Objective: in order to provide rapid and reliable method. Methods: Encoded Annexin Ⅴ cDNA was amplifyed from U937 cDNA libary by PCR and then subcloned into E coli expression vector. MS2-Annexin Ⅴ fusion protein could be overexpressed in E　coli. The MS2 bacteria protein could be removed by thrombin digestion.The mature Annexin Ⅴ was obtained by ion exchange chromatography and the FITC labled Annexin Ⅴ could be used in the detection of apoptosis. Results:Up to 37% of the total bacterial proteins was rhAnnexin Ⅴ as showed by SDS-PAGE. The purification of Annexin Ⅴ is over 99%. The FITC labled Annexin Ⅴ could efficiently detect apoptosis. Conclusion: We successfully established the technique procedure of obtaining a large quantity of Annexin Ⅴ and provided the basic routine for popularizing the detection of apoptosis' with high effciency.